Lab Anim Res.  2018 Mar;34(1):20-29. 10.5625/lar.2018.34.1.20.

Higher incidence of sperm granuloma in the epididymis of C57BL/6N mice

Affiliations
  • 1Department of Advanced Toxicology Research, Korea Institute of Toxicology (KIT), Daejeon, Korea. ybkim@kitox.re.kr
  • 2Department of Inhalation Research, Korea Institute of Toxicology (KIT), Jeongeup, Korea.
  • 3Department of Veterinary Pathology, College of Veterinary Medicine, Chungnam National University, Daejeon, Korea. hyson@cnu.ac.kr

Abstract

C57BL/6N mice are inbred strains widely used in biomedical research. Hence, a large amount of basic data has been accumulated. However, in the field of histopathology, spontaneous data for relatively younger mice that are used more frequently are not yet abundant, in contrast to data for older mice and their neoplastic lesions. To acquire the essential background data required by various research and toxicological assessments, 120 mice of the C57BL/6N strain (10 and 13 weeks of age) were collected from two institutions (From Korea and Japan) and subjected to histopathological analyses of the major organs (liver, spleen, kidney, thymus, heart, testis, epididymis). The results showed significantly higher incidence of sperm granulomas in the epididymides (10-56%) of these mice, compared with that in other strains or species of lab animals. Upon closer inspection, oligospermia/clear cell hyperplasia, cellular debris, and tubular vacuolation were also observed in the epididymides with sperm granulomas. Moreover, diseased organs were significantly heavier than healthy ones. Immunohistochemical staining showed a significant increase in the chromatic figures of cysteine-dependent aspartate-directed proteases-3 (caspase-3) and cleaved-poly(ADP-ribose) polymerase (c-PARP), and damages to the tubule due to spontaneous apoptosis, which may have led to the sperms leaking out of the tubule, causing the granuloma. To conclude, spontaneous sperm granuloma can occur in 10- and 13-week-old C57BL/6N mice and may thus affect the results of various studies using these mice. Therefore, sperm granuloma in epididymis needs to be carefully considered as an important factor when design the study using C57BL/6N.

Keyword

C57BL/6N; epididymis; sperm granuloma; histopathology; historical background; spontaneous lesion

MeSH Terms

Animals
Apoptosis
Epididymis*
Granuloma*
Heart
Hyperplasia
Incidence*
Kidney
Korea
Male
Mice*
Spermatozoa*
Spleen
Testis
Thymus Gland

Figure

  • Figure 1 Tail of the epididymis of 10-week-old C57BL/6N mice (G1 group). Sperm granulomas (arrows) are shown in the tail of the epididymis on the right, in contrast to the tail of a healthy epididymis on the left (a). Bar size; 2 mm. High magnification image showing a sperm granuloma in of 10-week-old mice; the sperms (S) leaking out of the tubules are visible at the center, together with granulomatous inflammation in the surrounding tissues. Sperms discharged into the interstitial space are being actively devoured by macrophages (arrowheads) (b). Bar size; 100 µm. High magnification image showing the epididymis body; the damaged tubule (arrows) from which the sperms leaked is indicted (c). Bar size; 200 µm.

  • Figure 2 High magnification image showing the changes associated with sperm granuloma in the epididymis of 10-week-old mice (G1 group). Absence of sperm in the lumen (L, oligospermia) and hyperplasia of clear cell (arrows) in epithelial cells (a). Macrovacuolation (arrowheads) in cytoplasm of epithelial cells (b). Bar size; 200 µm.

  • Figure 3 Absolute organ weights of the epididymis. Data are expressed as means±standard error of the mean (SEM) *: significant at the 0.01 level.

  • Figure 4 Image of the caspase-3 IHC staining in epididymis of lab animals. Significantly more caspase-3 chromatic figures are observed in the tubules of the C57BL/6N mouse epididymis (dotted line) than the other lab animal species, in which 1-2 chromatic figures are observed per tubule (arrows). C57BL/6N mouse (a), ICR mouse (b), BALB/c nude mouse (c), SD rat (d). Bar size; 400 µm.

  • Figure 5 Image of the SOD2 IHC staining in epididymis of lab animals. Positive SOD2 chromatic figures were observed in the tubules in tail of epididymis. C57BL/6N mouse (arrows) (a), ICR mouse (b), BALB/c nude mouse (c), SD rat (d). The chromatic figure was found in interstitial cells and in the internal cilia of the tubules. Bar size; 400 µm.

  • Figure 6 Results of c-PARP IHC staining for the area of the epididymis of C57BL/6N mice where the caspase-3 chromatic figure is found; caspase-3 (a), c-PARP (b). The staining of the serially sectioned area shows c-PARP chromatic figures (arrowheads) in the cellular nucleus of the tubules that exhibited caspase-3 chromatic figures (arrows). Bar size; 100 µm.


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